Caveolins and intracellular calcium regulation in human airway smooth muscle

Y.s. Prakash, Michael A. Thompson, Brianna Vaa, Ihaab Matabdin, Timothy E. Peterson, Tongrong He, Christina M Pabelick

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

Regulation of intracellular Ca2+ concentration ([Ca 2+]i) is a key factor in airway smooth muscle (ASM) tone. In vascular smooth muscle, specialized membrane microdomains (caveolae) expressing the scaffolding protein caveolin-1 are thought to facilitate cellular signal transduction. In human ASM cells, we tested the hypothesis that caveolae mediate Ca2+ responses to agonist stimulation. Fluorescence immunocytochemistry with confocal microscopy, as well as Western blot analysis, was used to determine that agonist receptors (M3 muscarinic, bradykinin, and histamine) and store-operated Ca2+ entry (SOCE)-regulatory mechanisms colocalize with caveolin-1. Although caveolin-2 coexpressed with caveolin-1, caveolin-3 was absent. In fura 2-loaded ASM cells, [Ca2+]i responses to 1 μM ACh, 10 μM histamine, and 10 nM bradykinin, as well as SOCE, were attenuated (each to a different extent) after disruption of caveolae by the cholesterol-chelating drug methyl-β-cyclodextrin. Transfection of ASM cells with 50 nM caveolin-1 small interfering RNA significantly weakened caveolin-1 expression and blunted [Ca2+]i responses to bradykinin and histamine, as well as SOCE, but the response to ACh was less intense. These results indicate that caveolae are present in ASM and that caveolin-1 contributes to regulation of [Ca2+]i responses to agonist.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume293
Issue number5
DOIs
StatePublished - Nov 2007

Fingerprint

Caveolins
Caveolin 1
Smooth Muscle
Caveolae
Calcium
Bradykinin
Caveolin 3
Histamine
Smooth Muscle Myocytes
Caveolin 2
Muscarinic M3 Receptors
Membrane Microdomains
Fura-2
Cyclodextrins
Vascular Smooth Muscle
Confocal Microscopy
Small Interfering RNA
Transfection
Signal Transduction
Fluorescence

Keywords

  • Intracellular signaling pathway
  • Methyl-β-cyclodextrin
  • RNA
  • Small interfering
  • Store-operated calcium entry

ASJC Scopus subject areas

  • Cell Biology
  • Physiology
  • Pulmonary and Respiratory Medicine

Cite this

Caveolins and intracellular calcium regulation in human airway smooth muscle. / Prakash, Y.s.; Thompson, Michael A.; Vaa, Brianna; Matabdin, Ihaab; Peterson, Timothy E.; He, Tongrong; Pabelick, Christina M.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 293, No. 5, 11.2007.

Research output: Contribution to journalArticle

Prakash, Y.s. ; Thompson, Michael A. ; Vaa, Brianna ; Matabdin, Ihaab ; Peterson, Timothy E. ; He, Tongrong ; Pabelick, Christina M. / Caveolins and intracellular calcium regulation in human airway smooth muscle. In: American Journal of Physiology - Lung Cellular and Molecular Physiology. 2007 ; Vol. 293, No. 5.
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